Expression of the relaxin-like factor (RLF) was studied at the messenger RNA (mRNA) and protein levels in the testes and ovaries of the mouse, as well as through testicular development and differentiation in the mouse testis. In situ hybridization or RT-PCR, and immunohistochemistry using a polyclonal antibody raised against a recombinant protein, provided mutually confirmatory results for a high expression of RLF in the Leydig cells of the adult testis and at a much lower level of expression in the luteal cells of the ovary through the cycle, pregnancy, and in lactation. Analysis of protein and mRNA expression, through postnatal testicular development, indicated moderate RLF expression also in the fetal population of Leydig cells, even in the hpg mutant mouse, lacking an active pituitary-gonadal axis. Prepubertal Leydig cells, however, exhibit only very low-level RLF gene expression, this phenotype persisting in the adult hpg mouse. In summary, fetal Leydig cells express RLF in an LH/human CG-independent fashion, whereas LH/human CG is essential to induce RLF expression in the adult-type Leydig cell. In cultured adult Leydig cells or in the mouse tumor MA-10 cell line, RLF mRNA is expressed in a constitutive fashion. RLF thus seems to be a useful marker of Leydig cell differentiation status.
SUMMARYThree ewes were intraduodenally infused with yeast RNA (15, 20 and 30 g/day) or control solutions for 3 days and the net changes in urinary allantoin excretion determined. Four mature wethers fitted with a re-entrant cannula in the proximal duodenum were fed two levels (0·66 and 1 × maintenance) of pelleted lucerne hay. Allantoin excretion was compared with total nucleic acid (NA) flow in the small intestine.The average proportion of the recovery of the infused RNA-N dose as urinary allantoin–N amounted to 0·119 ± 0·0046. The values of the net increase of allantoin-N (Y, g/day) above control levels were highly significantly correlated (n = 12, r = 0·98) with amounts of RNA-N infused (X, g/day): y = 0·1185 (± 0·0086) X–0·004 (P < 0·001). The average ratio of urinary allantoin-N to NA-N passing the duodenum during 24 h was 0·173. A difference between the two conversion factors is discussed in relation to endogenous allantoin excretion and purine base composition of yeast and bacterial RNA.
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